Evidence for epithelial-mesenchymal transition in cancer stem cells of head and neck squamous cell carcinoma.
Evidence for epithelial-mesenchymal transition in cancer stem cells of head and neck squamous cell carcinoma.
复制标题
DOI：
10.1371/journal.pone.0016466
复制
发表时间：
2011-01-27
期刊：
影响因子：
3.7
通讯作者：
Albers AE
中科院分区：
文献类型：
--
作者：
Chen C;Wei Y;Hummel M;Hoffmann TK;Gross M;Kaufmann AM;Albers AE
头颈部鳞状细胞癌（HNSCC）的发生、生长、复发和转移与肿瘤干细胞（CSC）的行为有关，肿瘤干细胞可通过其脱氢酶亚型-1（ALDH 1）活性鉴定。我们定量并富集了HNSCC细胞系中的ALDH 1+细胞，随后表征了其表型和功能特性，如侵袭能力和上皮-间充质转化（EMT）。球状体培养物富集来自五种HNSCC细胞系的CSC达5倍。通过流式细胞术和免疫荧光以及增殖和细胞周期分析，比较球体来源的细胞（SDC）和亲本单层来源的细胞系ALDH 1、CD 44、CD 24、E-Cadherin、α-SMA和波形蛋白的表达。通过Matrigel测定和实时PCR检测茎干相关转录因子（TF）Nanog、Oct 3/4、Sox 2和EMT相关基因Snail 1和2以及Twist的表达来评估侵袭活性。所有细胞系均形成可自我更新和连续再传代的球状体。ALDH 1在SDC中的表达显著升高。ALDH 1+细胞显示增加的集落形成。在单层和球状体培养物中，具有推定的CSC标志物CD 44 +/CD 24 −的细胞比例变化很大（0.5%-96%），与CD 44 +/CD 24 −/ALDH 1+细胞亚群重叠0%-33%。SDC具有显著较高的入侵活性。干细胞相关基因Sox 2、Nanog和Oct 3/4的mRNA在所有细胞系的SDC中显著增加。Twist在两个细胞系中显著增加，而Snail 2在一个细胞系中显示出显著增加，并且两个细胞系的SDC显著降低。SDC G 0期细胞比例较高，α-SMA和Vimentin高表达，E-Cadherin表达明显降低。HNSCC系具有潜在的CSC，其特征在于ALDH 1和干性标志物TF表达以及侵袭性、静止性和EMT等特性。非贴壁依赖性培养技术可以富集CSC，这对于研究CSC在肿瘤耐药、复发和转移中的作用具有重要意义。
Initiation, growth, recurrence, and metastasis of head and neck squamous cell carcinomas (HNSCC) have been related to the behavior of cancer stem cells (CSC) that can be identified by their aldehyde-dehydrogenase-isoform-1 (ALDH1) activity. We quantified and enriched ALDH1+ cells within HNSCC cell lines and subsequently characterized their phenotypical and functional properties like invasion capacity and epithelial-mesenchymal transition (EMT). Spheroid culture enriched CSC from five HNSCC cell lines by up to 5-fold. In spheroid-derived cells (SDC) and the parental monolayer-derived cell line ALDH1, CD44, CD24, E-Cadherin, α-SMA, and Vimentin expression was compared by flow-cytometry and immunofluorescence together with proliferation and cell cycle analysis. Invasion activity was evaluated by Matrigel assay and expression of stemness-related transcription factors (TF) Nanog, Oct3/4, Sox2 and EMT-related genes Snail1 and 2, and Twist by real-time PCR. All cell lines formed spheroids that could self-renew and be serially re-passaged. ALDH1 expression was significantly higher in SDC. ALDH1+ cells showed increased colony-formation. The proportion of cells with a putative CSC marker constellation of CD44+/CD24− was highly variable (0.5% to 96%) in monolayer and spheroid cultures and overlapped in 0%–33% with the CD44+/CD24−/ALDH1+ cell subset. SDC had significantly higher invading activity. mRNA of the stemness-related genes Sox2, Nanog, and Oct3/4 was significantly increased in SDC of all cell lines. Twist was significantly increased in two while Snail2 showed a significant increase in one and a significant decrease in SDC of two cell lines. SDC had a higher G0 phase proportion, showed high-level expression of α-SMA and Vimentin, but significantly decreased E-Cadherin expression. HNSCC-lines harbor potential CSC, characterized by ALDH1 and stemness marker TF expression as well as properties like invasiveness, quiescence, and EMT. CSC can be enriched by anchorage-independent culture techniques, which may be important for the investigation of their contribution to therapy resistance, tumor recurrence and metastasis.
登录
查看更多内容
影响因子：
11.2
作者：
Huang EH;Hynes MJ;Zhang T;Ginestier C;Dontu G;Appelman H;Fields JZ;Wicha MS;Boman BM
通讯作者：
Boman BM
影响因子：
64.8
作者：
O'Brien, Catherine A.;Pollett, Aaron;Dick, John E.
通讯作者：
Dick, John E.
影响因子：
5.2
作者：
Ghods, Ali Jourabcih;Irvin, Dwain;Yu, John S.
通讯作者：
Yu, John S.
影响因子：
3.7
作者：
Mack, Brigitte;Gires, Olivier
通讯作者：
Gires, Olivier
影响因子：
254.7
作者：
Parkin, DM;Bray, F;Pisani, P
通讯作者：
Pisani, P